Trigueiro-Louro, João M.Correia, VanessaSantos, Luís A.Guedes, Rita C.Brito, Rui M.M.Rebelo-de-Andrade, Helena2020-05-112020-05-112019-04-27Virology. 2019 Sep;535:297-307. doi: 10.1016/j.virol.2019.04.009. Epub 2019 Apr 270042-6822http://hdl.handle.net/10400.18/6649Influenza NS1 protein is among the most promising novel druggable anti-influenza target, based on its structure; multiple interactions; and global function in influenza replication and pathogenesis. Notwithstanding, drug development guidance based on NS1 structural biology is lacking. Here, we design a promising strategy directed to highly conserved druggable regions as a result of an exhaustive large-scale sequence analysis and structure characterization of NS1 protein across human-infecting influenza A subtypes, over the past 100 years. We have identified 3 druggable pockets and 8 new potential hot spot residues in the NS1 protein, not described before, additionally to other 16 sites previously identified, which represent attractive targets for pharmacological modulation. This study provides a rationale towards structure-function studies of NS1 druggable sites, which have the potential to accelerate the NS1 target validation. This research also contributes to a deeper comprehension and insight into the evolutionary dynamics of influenza A NS1 protein.Highlights: Anti-influenza strategies based on highly conserved target structures are needed; Overall, the human NS1 protein is highly conserved in the RBD and ED domains; Three main consensus druggable pockets were found with high druggability score; 8 new potential hot spots were identified within the NS1-ED; The study discloses a new panel for NS1 structure-function studies.engAntiviral AgentsBinding SitesComputational BiologyConserved SequenceDrug DevelopmentHumansInfluenza A virusProtein BindingProtein ConformationViral Nonstructural ProteinsDrug DesignResistência aos AntimicrobianosInfecções Respiratóriasjournal article10.1016/j.virol.2019.04.009